A volume of 4 μL of each MBP-GS fusion protein (20 μg/mL) was applied to the clean side of carbon on mica (carbon/mica interface). The carbon layer was subsequently floated onto a 2% sodium silicotungstate solution (pH 7.4), recovered with a 400 mesh copper grid (Agar) and air dried for 10 min. Micrographs were taken under low-dose conditions (exposing for 1 s at an electron dose of 30 e−/Å2) on a Philips CM12 microscope operated at 120 kV. Images were recorded at a nominal magnification of 22000X with a defocus of −1.5 μm on a Gatan Orius 1000 CCD camera, corresponding to a pixel size of 3.24 Å/pixel on the specimen.
Orius 1000 ccd camera
Manufactured by Ametek
Sourced in United States, Japan
The Orius 1000 CCD camera is a high-performance digital camera designed for scientific and industrial applications. It features a large-format CCD sensor and advanced image processing capabilities to capture detailed and high-resolution images. The camera is capable of capturing images with a resolution of up to 4096 x 4096 pixels, making it suitable for a wide range of applications that require precise image acquisition and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Jax 840a, by JEOL (6 mentions)
Nano z, by Malvern Panalytical (6 mentions)
Jem 1230, by JEOL (6 mentions)
Tecnai t12 lab6 em, by Thermo Fisher Scientific (2 mentions)
1011 transmission electron microscope, by JEOL (2 mentions)
Cm12 microscope, by Philips (1 mentions)
Tecnai 12 microscope, by Ametek (1 mentions)
1200 ex transmission electron microscope, by JEOL (1 mentions)
Tecnai 12 lab6 electron microscope, by Ametek (1 mentions)
Dynapro nanostar instrument, by Wyatt Technology (1 mentions)
Filter paper, by Cytiva (1 mentions)
Digital micrograph software, by Ametek (1 mentions)
Em pact2, by Leica camera (1 mentions)
Csu x1 confocal scanner, by Yokogawa (1 mentions)
Ultracut uct, by Leica camera (1 mentions)
Cm12 transmission electron microscope, by Philips (1 mentions)
Ags160, by Agar Scientific (1 mentions)
Cacodylate buffer, by Electron Microscopy Sciences (1 mentions)
Cm 12 transmission electron microscope, by Ametek (1 mentions)
K3fe cn 6, by Merck Group (1 mentions)
30 protocols using orius 1000 ccd camera
1
MBP-GS Fusion Protein Visualization
2
Negative-Staining EM for Purity Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Negative-staining EM allowed purity and concentration assessment, as well as adapting the T5 tail:FhuA ratio prior to cryo-grid preparation. Negative-stain grids of all samples were prepared using the mica-carbon flotation technique over a 2% sodium silicotungstate solution and were observed with an FEI/Thermo Fisher Scientific Tecnai 12 microscope (LaB6 electron source, 120 kV) equipped with a Gatan Orius 1000 CCD camera.
3
Negative-Stain Electron Microscopy of Protein Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein samples were visualized by negative-stain electron microscopy (EM) using 3-4 μL aliquots containing 0.1-0.2 mg/ml of protein. Samples were applied for 10 s onto a mica carbon film and transferred to 400-mesh Cu grids that had been glow discharged at 20 mA for 30 s and then negatively stained with 2% (wt/vol) Uranyl Acetate (UAc) for 30 s. Data were collected on a FEI Tecnai T12 LaB6-EM operating at 120 kV accelerating voltage at 23k magnification (pixel size of 2.8 Å) using a Gatan Orius 1000 CCD Camera. Two-dimensional (2D) class averaging was performed with the software Relion122 (link) using on average 30–40 micrographs per sample. The 5 best obtained classes were calculated from around 6000 particles each.
4
Transmission Electron Microscopy Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Electron microscopy was performed as previously described 10 , 23 . Cells grown on carbon‐coated sapphire disks were cryoprotected with DMEM containing 10% FCS and frozen at high pressure (HPM 10 Abra Fluid AG). Samples were then freeze‐substituted and embedded in lowicryl HM20. Thin sections were collected on formvar‐/carbon‐coated nickel slot grids and stained with uranyl acetate and lead citrate. Imaging was performed with a transmission electron microscope (Philips CM12) coupled to an Orius 1000 CCD camera (Gatan). Quantification by stereology was performed as previously described 10 , 23 .
5
Electron Microscopy of Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HUVECs were cultured in a 24-wells plate. GYY4137, zinc, and GYY4137 together with zinc were added, and after 30 min incubation, cells were embedded. HUVECs were fixed with 2%/2% formaldehyde/glutaraldehyde for 30 min at room temperature. Cells were washed 3 × in 0.1 M PBS. A second fixation was performed in 1% osmium tetroxide in 0.1 M PBS for 1 h at 4 °C. After washing (3 times in water), cells were gradually dehydrated in graded series of ethanol and then gradually infiltrated with resin at room temp. Gelatin capsules were put upside-down on the coverslips and polymerised for 24 h at 60 °C. Samples were lightly heated to remove the glass coverslip and cut on an ultramicrotome Reichert S at 90 nm of thickness. Acquisitions were performed on a JEOL 1011 TEM with a Gatan Orius 1000 CCD Camera.
6
Characterization of HBEON Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Scanning and transmission electron micrographs (TEM and SEM) were done using JEOL JAX-840A and JEOL JEM- 1230 electron micro-analyzers for HBEON. The acquisition of the image was done by Orius 1000 CCD camera (GATAN, Warrendale, PA, USA). HBEON sample was sonicated (30–60 min) immediately to prevent the coalescence of the nanoparticles before the assessment of zeta potential. The average diameter of the synthesized nanoparticles was calculated by zpw 388 version 2.14 nicomp software. However, the zeta potential and size distribution were done by a particle size analyzer (Nano-ZS, Malvern Instruments Ltd., UK).
7
Ultrastructural Analysis of Cerebellum in Mouse Models
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
P21 En1Cre; Srd5a3fl/- (n = 1), Atoh1Cre; Srd5a3fl/- (n = 2) and control littermates (n = 3) were perfused with 4% PFA/2% glutaraldehyde. The cerebellum was kept in the same solution for at least one week. Sagittal slices, less than 1 mm thick, were post-fixed with 1% osmium tetroxide in 0.1 M phosphate buffer and then dehydrated in ethanol. After 10 min in a 1:2 mixture of epoxy propane and epoxy resin and 10 min in epon, samples were covered by upside down gelatin capsules filled with freshly prepared epoxy resin and polymerized at 60°C for 24 hr. After heat separation, ultrathin sections of 90 nm were cut with an ultra-microtome (Reichert ultracut S), stained with uranyl acetate and Reynold’s lead and observed with a transmission electron microscope (JEOL 1011). Acquisition was performed with a Gatan Orius 1000 CCD camera.
8
Transmission Electron Microscopy Sample Preparation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were fixed with paraformaldehyde 2% and glutaraldehyde 3.75%. After several thorough washes in PBS buffer, the cells were post-fixed in 1% osmium tetroxide and embedded in epoxy resin. Semithin sections of 300–400 nm were first realized to control the samples and 80–90 nm sections were then cut on a Reichert Ultracut E ultramicrotome. These ultrathin sections were next transferred onto 200-mesh copper grids prior to staining with uranyl acetate and lead citrate. The sections were then viewed under a JEOL 1011 transmission electron microscope with a GATAN Orius 1000 CCD camera.
9
Characterization of ZnO-NPs and Encapsulated Thyme Oil
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transmission electron micrographs (TEMs) were recorded on JEOL JAX-840A and JEOL JEM-1230 electron micro-analyzers, respectively. The ZnO-NP samples were scattered in ethanol and then treated ultrasonically to disperse the individual particle over the gold grids (Mahamuni et al. 2019 (link)). However, for encapsulated thyme oil (ETO), the droplets were placed onto a carbon-coated copper grid to form a thin liquid film and were negatively stained by one drop of uranyl acetate. The excess of staining was removed using filter paper then the film was air-dried before the observation (Pecarski et al. 2014 (link)). The Orius 1000 CCD camera was used for image acquisition (GATAN, Warrendale, PA, USA). For measuring zeta potential, the sample of ZnO-NPs or ETO was sonicated for 30–60 min just before assessment. The average diameter was calculated using zpw 388 version 2.14 nicomp software. The size distribution and zeta potential of ZnO-NPs and ETO were measured using a particle size analyzer (Nano-ZS, Malvern Instruments Ltd., UK).
10
Immunogold Labeling Optimization for TEM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
All thin sections were imaged in a CM 100 transmission electron microscope (FEI, Eindhoven, The Netherlands) at an acceleration voltage of 80 kV using a Gatan Orius 1000 CCD camera (Gatan, Munich, Germany). For immunogold-labelled sections, it can be difficult to distinguish cellular components like ribosomes, which may stain heavily with heavy metals and can look like gold particles. This can be resolved by varying the histogram on the live view mode of the camera to make clearer the difference in contrast. Membrane delimitations of membrane-bound structures can be difficult to resolve on one section only, due to the dependence on the angle at which the section cuts through the structures. By analysing consecutive serial sections, this problem can be easily resolved as some perspective on the 3D structure can be obtained. Using this approach, we could distinguish lipid droplets from vacuoles in HPF and PF sections.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!